Authors: Leire Bergara-Muguruza, Heba E. Abdel Razik, Peng Chen, Maki Ashida, Hanawa Takao, Pekka Vallitu, Miho Nakamura
Abstract: Calcium carbonate (CC)-hydroxyapatite (HAP) porous microparticles have gained a lot of popularity as a promising material for clinical applications. The objective of this study is to evaluate the effects of CC-HAP microparticles on osteoblast-like cells to be used as a bone-regeneration biomaterial. In this study, the different concentrations of conditioned media were used to compare the effects of released ions from CC-HAP microparticles. The material’s characteristics demonstrated that the immersion in cell culture medium did not change the crystal phases of CC-HAP. The decrease of calcium ions in cell culture medium is due to the dissolution-precipitation reactions on the material surfaces, which made more crystalline surfaces. The atomic absorption spectroscopy measurement demonstrated that the dissolution-precipitation reactions on the material surfaces in cell culture medium happened in 3 days and were stable between 3 to 5 days. The conditioned media immersed in cell culture medium for 4 days were used for further experiments. Cell evaluations demonstrated that excessive adding of CC -HAP could inhibit cell behaviors such as cell adhesion, proliferation, and differentiation. The cell adhesion indicated by the number of vinculin-positive focal adhesions per cell decreased with the increase of the CC-HAP concentrations. The cells cultured with CC-HAP proliferated at a lower rate than the control without CC-HAP. One of the reasons for the inhibition of cell proliferation was thought to be less formation of focal adhesions with higher concentrations of CC-HAP. The excessive adding of CC-HAP had an inhibitory effect on osteoblast differentiation. The results of this study revealed that the conditioned media prepared by immersion of CC-HAP porous microparticles in cell culture media had effects on the behaviors of osteoblast-like cells such as cell adhesion, proliferation, and differentiation.
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Authors: Thet Thet Swe, Hasmaliza Mohamad, Khairul Anuar Shariff, Kunio Ishikawa
Abstract: This study concerns the evaluation of the bioactivity and cells response of strontium (Sr) doped sol-gel derived S53P4 bioglass due to Sr induced osteoblast. Moreover it prevents in-vitro osteoclastic activity and is clinically used as osteoporosis treatment. The different amount of Sr was doped into the S53P4 bioglass formulation (53.82%SiO2-1.72%P2O5-22.64%Na2O-(21.76-x)%CaO-x%SrO) (x=0, 3 and 5 mol %) and synthesized via sol-gel method. These samples were denoted as 0Sr, 3Sr and 5Sr respectively. After soaking in Hank's balanced salt solution (HBSS) for 7 and 14 days, the apatite formation was examined using X-ray powder diffraction (XRD) and scanning electron microscope (SEM) techniques. Proliferation and alkaline phosphatase activity were evaluated using osteoblastic cell line MC3T3-E1. The XRD and SEM findings confirmed the hydroxyapatite (HA) structure on the bioglass surface after soaking. More intense HA peaks were observed in 3Sr specimen on 7 day while in 5Sr specimen on 14 day. Meanwhile, 3Sr specimen showed the highest cells proliferation and significant difference in alkaline phosphatase (ALP) activity than 0Sr and 5Sr. As a result, this finding indicates that S53P4 bioglass with 3 mol % SrO (3Sr) is a good candidate for bone tissue engineering because it allows for optimum cell proliferation and ALP activity while also having a high bioactivity efficiency.
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Authors: Niyou Wang, S. Thameem Dheen, Jerry Ying Hsi Fuh, A. Senthil Kumar
Abstract: Daily walking or exercise of the bone implant recipients may generate particles due to wear and tear. Reports have mentioned that particles could circulate in the human body and trigger aseptic loosening, inflammation, and other potential complications. The mechanism of these phenomena remains mostly unclear. This study is to investigate the cytotoxicity of titanium (Ti), stainless steel 316 (SS316), and magnesium (Mg) particles due to these materials are the most commonly used biomaterials based on their adequate mechanical properties and excellent biocompatibility. Human osteoblasts (SAOS2 cells) were exposed directly to different concentrations of Ti/SS316/Mg particle during the direct cytotoxicity test. Together with the previous study, we found out that Ti particles showed cytotoxicity to osteoblasts at different dosages and times, while SS316 particles and Mg particles (low dosage) can reduce the cytotoxicity induced by Ti particles and boost cell viability. Mg particles can be toxic to osteoblast at a higher dosage, while SS316 particles are “safer” than Mg particles at higher dosages. Cell viability and cell morphology of SAOS2 cells under different treatments were observed at 2/3/5 days. This study found out that cell viability could be enhanced with certain combinations of Ti/SS316/Mg particles. This can give us certain guideline on how to design and fabricate a hybrid bone implant. However, how to quantify the particles inside the human body in real-time, and the exact interaction among particles, cells, tissues, and even organs require further research.
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Authors: Edrizal Edrizal, Abidin Trimurni, Deddi Prima Putra
Abstract: Punica granatum (PG) contained anthocyanin, the chemical compound that played significant role in increasing bone cell proliferabbitsion and osteoblasts differentiation in bone remodelling. This research was aimed at measuring the effect of Punica granatum on maxilla bone remodelling relating to the profile of both osteoblast and osteoclast. Punica granatum was extracted by using butanol while the analysis of bone remodelling was conducted by observing the profile of osteoblast and osteoclast cells using histopathologic method. Butanol hydrogel of Punica granatum was abled to enhance bone reabsorption which was proven by the increase of osteoclast and new bone formation since the osteoblast was presented. Moreover, bone remodelling activity was characterised by increased collagen fibres and osteocyte cell, in addition, bone shape compactness was getting better. Statistical analysis revealed that the histoscore profile of osteoblast was influenced by butanol fraction dosage with significance level (p<0.05) which was at medium criteria according to Pearson correlation (r=0,6). While the effect of dosage on osteoclast was at significance level (p<0.05) but was poorly correlated with (r=0.01) and osteoblast frequency was dominant at the concentrabbitsion of 125 mg/ml and 250 mg/ml. On the other hand, at the concentrabbitsion of 175 mg/ml osteoclast presentation was found to be more dominant. The fraction of butanol PG was abled to induce osteoblast increment and osteoclast at the concentrabbitsion of (mg/ml) 125, 175, and 250 and accelerabbitse the remodelling of alveolar maxilla.
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Authors: Kenta Nisogi, Satoshi Okano, Sengo Kobayashi, Kensuke Kuroda, Takeaki Okamoto
Abstract: Surface wettability is thought to influence the osteoconductivity of bone-substituting materials; however, the effects of surface wettability on osteoblast behavior are not well understood. In this study, we prepared both an as-polished pure titanium with a water contact angle (WCA) of 57° and heat-treated pure titanium with more hydrophobic surface and WCAs of 68°-98°. The effects of the surface wettability of pure titanium on osteoblast behaviors were evaluated by in vitro assays. Compared with the as-polished titanium, the proliferation rate of osteoblast increased on heat-treated titanium. This suggested that surface wettability affects osteoblast behaviors, meaning osteoconductivity is influenced by surface wettability.
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Authors: Supatsara Rujanant, Woo Kul Lee, Sasithorn Kongruang
Abstract: In tissue engineering, biomaterials used for bone tissue substitutes attract increasing interests, especially for finding biologically active compounds that can activate proliferation of osteoblastic MG63 cells. The evaluation of the impact of a soluble yeast-derived β-(1-3), (1-6)-D-glucan (BG) extracted from distillery waste yeast sludge on viability and proliferation of MG63 cells was studied. Spray dried BG prepared from alkaline extraction was used as supplementary activator in osteoblastic cell culture system. The composition of BG was characterized using FTIR spectral analysis and BG analysis assay kit. MG63 human osteoblast cell-line was cultured on Dulbecco’s modified’s medium supplemented with various concentrations of BG ranging from 0.1 to 1.0 mg/mL. The cells were cultured up to 7 days under a humidified 5% CO2 atmosphere at 37°C and monitored the level of proliferation at pre-determined intervals. Results showed that increase in BG concentration substantially promoted MG63 cell proliferation. Optimal concentration was identified and found at 0.3 - 0.7 mg/mL. Results revealed that BG could be further utilized for the upregulation of osteoblastic proliferation positively related to the acceleration of bone regeneration.
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Authors: Eui Ri Na, Jong Wook Moon, Young Joon Kim
Abstract: Perforations can occur during endodontic treatment, post placement and removal, and operative procedures. These defects have been treated with a variety of different materials such as resin ionomer, glass ionomer cement and intermediate restorative material. However, the osteogenic response to these substances using osteoblasts have been rarely studied. Thus, the aim of the present study is to evaluate the osteogenic response to resin ionomer (Geristore) and mineral trioxide aggregate (MTA). The surface roughness was significantly higher in the MTA than in the resin ionomer (p<0.05). After 72 hours of incubation mouse osteoblasts attached and spread well over the surfaces of resin ionomer and MTA. As a result from MTT assay, the number of cells gradually increased as the cell incubation time increased. In particular, control group showed higher cell proliferation than the other two groups on days 3 and 5. Resin ionomer showed more active early cell proliferation than MTA (p<0.05). The alkaline phosphatase (ALP) activity was significantly higher in the MTA surface than in the resin ionomer and glass coverslip (p<0.05). Resin ionomer was active in early cell proliferation and adhesion. Resin ionomer may be more suitable for cervical perforation or for perforation of adjacent to the gingiva requiring rapid wound closure. Also, MTA has a rough surface and low initial cell adhesion but because of its superior osteogenic response, it may be appropriate for the area close to the apical region, where the perforation site is wide and the bone tissue regeneration is necessary.
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Authors: Michiyo Honda, Mamoru Aizawa
Abstract: Vascularization is a crucial process during bone development and regeneration. A number of studies have shown that the interaction between osteoblasts and endothelial cells plays a key role in osteogenesis by using co-culture system. However, vascularization strategies in cell-based bone tissue engineering depend on optimal culture conditions. In this study, we determined the optimal co-culture conditions in view of osteogenic parameters and examined the effects of angiogenic properties on osteogenesis. As for cell proliferation, the proportion of osteoblasts increased and that of endothelial cells decreased as culture period passed. Assessment of osteogenic differentiation shows that co-culture of osteoblasts and endothelial cells significantly increased alkaline phosphatase activity and expression of bone-related genes. Furthermore, abundant microcapillary-like structures were observed which endothelial cells self-assembled into branches and net-like structures. The use of endothelial cells would be a promising strategy to promote vascularization to support the bone regeneration. Combination of these cell-based approaches and tissue engineering like three-dimensional scaffolds could provide a novel treatment therapy for bone defects and bone diseases.
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Authors: Masanobu Kamitakahara, Takashi Shirato, Taishi Yokoi, Hideaki Matsubara, Yasuaki Shibata, Tohru Ikeda
Abstract: Silicate-containing alpha-tricalcium phosphate (α-TCP) ceramics are expected to be useful scaffolds for bone regeneration because α-TCP shows high biodegradability and silicate ions are expected to promote the bone formation. We previously revealed that the porous silicate-containing α-TCP granules provided earlier bone formation and showed lower biodegradability than the porous silicate-free α-TCP granules in vivo. In order to reveal the mechanism of the bone formation promoted by silicate incorporation, the proliferation and differentiation of osteoblast-like cells on the silicate-containing and silicate-free α-TCP ceramics were examined in vitro. The silicate incorporation in α-TCP promoted the differentiation of osteoblast-like cells, and it might be one of the factors to promote bone formation In Vivo.
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Authors: S.B. Cho, G.J. Yoon, E.M. An, Y.J. Kim, T.N. Kim, H.D. Jang, J.Y. Choi, I. Noh
Abstract: We reported the apatite-forming ability of 30CaO∙70SiO2 scaffolds with 0~100 ppm Ag ions by soaking in simulated body fluid (SBF). This study was to evaluate the effects of the concentrations of Ag ions in the 30CaO∙70SiO2 gels on in-vitro biocompatibility of osteoblasts (MC3T3). After seeding cells on the surface of Ag-30CaO∙70SiO2 gels scaffold, cellular behaviors were evaluated by an assay of cell counting kit-8. Cytotoxicity of the scaffold samples was evaluated by employing the extract solutions of the scaffold samples by the assays of neutral red, MTT and BrdU. In addition, live & dead assay was performed by using a gel covering method, which the scaffolds have been directly contacted with the incubated cells on the well plate. According to the results of CCK-8 assay, the optical density value of the absorbance of the resulting solution decreased as the concentration of Ag ions in the scaffolds increased. Moreover, their cell viability was measured to be less than 50% at the Ag concentrations of 50 ppm or more, and dead cells were observed in the experiment results of both the cytotoxicity and gel covering tests. From these experimental results, we concluded that the Ag-30CaO∙70SiO2 scaffolds with less than 50 ppm Ag ion concentration were considered as biocompatible.
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